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Merck

Generation of TCRα-transduced T cells for adoptive transfer therapy of salmonellosis in mice.

STAR protocols (2021-03-23)
Anastasiia Kalinina, Alexandra Bruter, Ludmila Nesterenko, Ludmila Khromykh, Dmitry Kazansky
RESUMEN

Adoptive transfer therapy has great potential to treat diseases such as cancer as well as autoimmune and infectious diseases. Identification of chain-centric T cell receptors (TCRs) with the dominant-active antigen-specific α-chains (TCRα) can significantly improve the efficacy of adoptive cell therapy while reducing time, labor, and costs of generation of TCR-modified antigen-specific T cells. This protocol describes how to generate salmonella-specific TCRα-modified mouse T cells by retroviral transduction and evaluate their functional activity in vivo in the mouse model of salmonellosis. For complete details on the use and execution of this protocol, please refer to Kalinina et al. (2020).

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Sigma-Aldrich
Concanavalin A from Canavalia ensiformis (Jack bean), Type IV-S, lyophilized powder, aseptically processed, BioReagent, suitable for cell culture
Sigma-Aldrich
Hexadimethrine bromide, ≥95%
Roche
Interleukin-2, mouse (mIL-2), recombinant (E. coli)